Toxic Effects Of Ibuprofen And Triclosan On P450Enzymes And Antioxidative System Of The Yellow Catfish(Pelteobagrus Fulvidraco)

Pharmaceuticals and personal care products （PPCPs） are a diverse group ofpotential increasing attention environmental pollutants which include manystructurally and functionally varieties of chemical compounds such as antibiotics,nonsteroid anti-inflamatory drugs, cardiovascular drugs and UV-filters etc. PPCPs arewidely used in many fields like medicine, hygiene and agriculture-farming activity etc.Therefore, they are likely to be found in environmental compartments including anywater borne, sediment and biota. Most PPCPs have not any acute toxic effects onnon-target organisms in environmental relevant concentrations, but chronic toxiceffects on metabolic systems of nontarget organisms under the exposure of lowconcentrations of PPCPs have not been assessed fully. In the present study, the toxiceffects of Ibuprofen and Triclosan on the phaseＩmetabolic enzymes and antioxidantdefence system of the yellow catfish （Pelteobagrus fulvidraco） were investigated.The toxic effects of environmental relevant level ibuprofen via waterborneexposure showed that APND, ERND, EROD and GST enzymes activity were inducedat the initial stages and gradually inhibited with the extension of exposure time. Thehighest value ocurred at24h for GST and at168h for CAT. The content of MDAdecreased significantly after24h exposure and come to the highest value at72h.ERND and MDA was the more sensitive measured parameters and may be used aspotential biomarkers for IBU exposure.The chronic toxicity of TCS (0.5,5,50and500μg.L^-1) to P. fulvidraco indicatedthat the highest value of APND ocurred at168h. The actvity of EROD was inhibited,only the lowest concentration (0.5μg.L^-1) at72h significantly induced. ERND wasinduced generally. The antioxidant metabolism enzymes of GST, CAT and MDAcontent showed different changing trends. EROD and GST were more sensitivemeasured parameters and may be used as potential biomarkers for TCS exposure.When P. fulvidraco were exposed to the mixtures of IBU and TCS, APND andEROD were inhibited at the initial stages then gradually induced with the extension of exposure time. But the activities were inhibited at168h again. ERND enzymesactivity were inhibited at the initial stages and gradually returned to the normal levelwith the extension of exposure time. GST activity was inhibited at24h and168h,no obvious change at72h. The activity of CAT inhibited significantly after24hexposure and come to a higher value at72h. EROD, CAT and MDA were sensitiveand suitable to be used as potential biomarkers for the mixture exposure of IBU andTCS compared with the other parameters.Primers were designed using the conserved domains after multiple alignmentsof previously reported full-length cDNA sequences of teleost CYP3A from theGenBank. The full-length cDNAs of P. fulvidraco CYP3A gene were obtained usingRT-PCR and RACE PCR amplification. The result of sequence analysis indicatedthat the full length of P. fulvidraco CYP3A cDNA was1737bp, containing an openreading frame （ORF） of1440bp, encoding479amino acids of putative protein.Alignment of P. fulvidraco CYP3A amino acid sequence with other species and onthe phylogenetic with those of other species tree, P. fulvidraco CYP3A gene wasclustered with the fish of cypriniformes. Search results of the BLAST databaserevealed that the deduced amino acid sequence was highly homologous withGobiocypris rarus, shared79%of amino acid. And the homologous with otherfishes was greater than65%, nearly55%of homology with mammals. The resultsshowed that the CYP3A gene of P. fulvidraco was successfully cloned.The full-length cDNAs of P. fulvidraco GST gene were obtained usingRT-PCR and RACE PCR amplification. The result of sequence analysis indicatedthat the full length of P. fulvidraco GST cDNA was1103bp, containing an openreading frame （ORF） of672bp, encoding223amino acids of protein. Alignment ofP. fulvidraco GST amino acid sequence with other species and on the phylogeneticwith those of other species tree, P. fulvidraco GST gene was clustered with the fishof Ictalurus punctatus. Search results of the BLAST database revealed that thededuced amino acid sequence was highly homologous with I. punctatus, shared90%.And the homologous with other fishes was greater than60%. The results showed that the GST gene of P. fulvidraco was successfully cloned.